Chronic Injury of Mice Bone Marrow Multipotent Hematopoietic Progenitor Cells Induced by Ionizing Radiation / 中国实验血液学杂志

OBJECTIVE@#To explore the chronic injury and its possible mechanism of ionizing radiation on multipotent hematopoietic progenitor cells (MPPs) by determining the related indicators of MPPs in bone marrow of mice post-radiation.@*METHODS@#Sixteen C57BL/6 adult mice were randomly divided into normal control and irradiation groups, 8 mice in each group. The mice in irradiation group were exposed to 6 Gy X-ray. The proportion of bone marrow MPPs, their apoptosis and proliferation 2 months after irradiation were detected by flow cytometry. Mitochondrial activity and levels of reactive oxygen species (ROS) in each MPPs population were detected by Mitotracker Red and DCFDA probes, and the senescent state of MPPs in the bone marrow was analyzed.@*RESULTS@#Ionizing radiation could reduce the proportion of MPPs in mouse bone marrow. The proportions and numbers of MPP1, MPP3 and MPP4 in the bone marrow were significantly decreased after whole-body irradiation with 6 Gy X-ray (P<0.05). In addition, radiation significantly reduced the colony-forming capacity of MPPs in bone marrow (P<0.05), the proportions of apoptotic cells in the MPP1 and MPP4 cell populations increased significantly in the bone marrow (P<0.05). The activity of mitochondria was significantly reduced in the bone marrow MPP2, MPP3 and MPP4 cell populations compared with that of the control group (P<0.05). It was also found that the radiation could significantly increase the ROS levels of MPPs in bone marrow, and the content of ROS in the MPP2, MPP3 and MPP4 cell population of the bone marrow was significantly increased(P<0.05). The senescent cells ratios of MPP1, MPP3 and MPP4 cells in the bone marrow after irradiation were significantly higher than those in the control group (P<0.05).@*CONCLUSION@#Ionizing radiation can cause chronic MPPs damage in mice, which is closely associated with persistent oxidative stress, cells apoptosis, and cellular senescence.

CD34+ hematopoietic progenitor cells enumeration and mononuclear cell count: an experience from a tertiary care centre

Background: This study was conducted to assess the CD34+ hematopoietic progenitor cells enumeration by flow cytometry and the utility of performing mononuclear cell count before performing the Stem cell enumeration. Collection was done on two consecutive days with CD34+ hematopoietic progenitor cell enumeration of both the samples. Mononuclear cell counts were done in all the patients. The purpose of the study was to do counts directly from the leukapheresis pack and see the reliability of this practice.Methods: Samples were collected from the leukapheresis pack and subjected to mononuclear cell count and CD34+ hematopoietic progenitor cells enumeration by flow cytometty before harvesting.Results: A total of 66 samples from 34 patients were taken up for the study. 76.47% of our cases were that of multiple myeloma and 17.64% of the cases were that of non Hodgkin lymphoma and 2.94% cases each of neuroblastoma and Hodgkin lymphoma. It was noted that the mononuclear cell counts correlated well with the CD34+ HPC in most of the cases with MNC being above 4 x 108 per pack per kg body weight in cases where CD34+ HPC counts were more than the desired lower limit of 2 x 106 per pack per kg body weight.Conclusions: It was observed that flow cytometric enumeration of CD34+ hematopoietic progenitor cells directly from the leukapheresis pack gave satisfactory results even without doing peripheral blood CD34+ HPCs enumeration before leukapheresis. Also, in our study we were able to set a limit of mononuclear Cell at 4 x 108 per pack/kg BW as counts beyond that always correlated with the more accurate flow cytometric method of CD34+ HPC count of more than 2 x106 per pack/kg body weight, therefore acting as a crude method for assessing the mobilization.

Expression of adhesion molecules on CD34+ cells from steady-state bone marrow before and after mobilization and their association with the yield of CD34+ cells

BACKGROUND: Cell adhesion molecules (CAMs) expressed on hematopoietic progenitor cells (HPCs), endothelial cells, and stromal cells play a pivotal role in the mobilization of CD34+ cells. Herein, we conducted a non-randomized peripheral blood stem cell (PBSC) mobilization study aimed to compare the potential differences in the expressions of several CAMs and chemokines on CD34+ cells obtained from bone marrow aspirate before and after HPC mobilization from patients with hematologic malignancies and healthy donors. METHODS: Three-color cytofluorometric analysis was used to compare the expressions of CAMs and chemokines in the bone marrow before and after mobilization. RESULTS: For all studied groups, CAM expression among those with good and poor yields of CD34+ cells was significantly correlated with VCAM-1 (P=0.007), CD44 (P=0.027), and VLA-4 (P=0.014) expressions. VCAM-1 (P=0.001), FLT-3 (P=0.001), CD44 (P=0.011), VLA-4 (P=0.001), and LFA-1 (P=0.001) expressions were higher before HPC mobilization than after HPC mobilization. By contrast, the expression of CXCR4 significantly varied before and after mobilization only among those with successful PBSC mobilization (P=0.002). CONCLUSION: We attempted to identify particular aspects of CAMs involved in CD34+ cell mobilization, which is a highly complex mechanism that involves adhesion molecules and matrix metalloproteases. The mechanism by which CD34+ cell mobilization is activated through proteolytic enzymes is not fully understood. We believe that CXCR4, VLA-4, CD44, and VCAM-1 are the most important molecules implicated in HPC mobilization, particularly because they show a correlation with the yield of CD34+ cells collected via large volume leukapheresis.

The effect and mechanism of indole-3-carbinol on alleviating the radiation-induced bone marrow hematopoietic cell injury in mice / 天津医药

Objective To investigate the protective effect of indole-3-carbinol (I3C) on radiation-induced mouse bone marrow hematopoietic cell injury and the involved mechanisms. Methods (1) The bone marrow nuclear cells (BMNCs) from CD45.1 subtype of C57BL/6J mice were collected by a density gradient centrifugation method. The BMNCs were pretreated with a series doses of I3C (0 mol/L, 10-8 mol/L-10-3 mol/L) and then exposed with radiation of 137Csγ-ray (doses of irradiation were 0 Gy, 1 Gy and 4 Gy). After 18-hour culturing, the bioluminescence method was used to detect the cell viability. (2) These cells were divided into control group and 10-6 mol/L I3C group. Both groups were received the irradiation (0 Gy, 1 Gy and 4 Gy) and inoculated into the methylcellulose semi-solid culture medium to incubate 7 days, the colony forming unit-granulocyte monocytes (CFU-GM) were observed. (3) Twenty-four CD45.2 subtype mice used as the receptor were exposed with 8 Gy radiation. The CD45.1 BMNCs were divided into control group, 4 Gy irradiation group, 4 Gy irradiation and 10-6 mol/L I3C group. Donor cells were harvested from C57BL/6J (CD45.1) mice after they received various treatments, and were then mixed with competitive BMNCs from C57BL/6J (CD45.2) mice. The mixed cells were transplanted into recipient mice (8 mice/group). Flow cytometry was used to analyze the proportion of donor cells in peripheral blood of receptor. (4) The cells were divided into control group, 10-6 mol/L I3C group, 1 Gy irradiation group, 1 Gy irradiation with 10-6 mol/L I3C group. After 24-hour culturing, Western blot assay was used to detect the expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2) and hemeoxygenase-1 (HO-1). Results (1) I3C showed a significant cytotoxic effect on the BMNCs when its concentration was above 10-4 mol/L. 10-7-10-6 mol/L I3C could reduce the radiation injury of BMNCs under the same dose of irradiation. Therefore, 10-6 mol/L I3C was chosen for subsequent experiments. (2) The CFU-GM was significantly higher in 10-6 mol/L I3C group than that of control group (P<0.05). (3) Results of flow cytometry showed that the proportion of donor cells in receptor was significantly higher in 4 Gy irradiation group than that of control group, which decreased the engraftment capability of irradiated HSCs (P<0.05), although the engraftment capability of irradiated HSCs improved after 10-6 mol/L I3C treatment. (4) I3C significantly enhanced the increased protein expression of Nrf2 and HO-1 caused by radiation (P<0.05). Conclusion I3C has a protective effect on hematopoietic cells following radiation-induced injury, which may be related with activating the Nrf2/HO-1 signal pathway.

Effect of different doses of HS6101 on hematopoietic function of ICR mice treated with cyclophosphamide / 国际药学研究杂志

Objective: To observe the effect of different doses of HS6101 on the recovery of hematopoietic injury in ICR mice treated with cyclophosphamideCTX. Methods: Normal ICR mice were intraperitoneally injected with CTX at 100 mg/kg once a day for 3 consecutive days， and the mouse model of chemotherapy-induced hematopoietic injury was established. Three groups of mice with 20 per group， were respectively injected with HS6101 at 0， 9 or 27 μg subcutaneously at one hour before the first administration of CTX. The peripheral blood cell counts of the mice were observed once every 2 days. Hematopoietic progenitor cell colony counting and histopathological assessment of bone marrow cells were evaluated at 4 d and 9 d after the first administration of CTX. Results: In ICR mice after chemotherapy with CTX， all doses of HS6101 significantly increased peripheral leukocytes and neutrophils P<0.05， elevated the number of multilineage hematopoietic progenitor cell colonies of bone marrow， and stimulated the proliferation of bone marrow cells after CTX injury. Mice receiving 27 μg HS6101 were better than those of the other two groups. Conclusion: HS6101 at 27 μg could significantly promote the recovery of hematopoiesis in ICR mice treated with CTX chemotherapy.

Effect of Bu-Shen Yi-Sui Sheng-Xue Method for Medicated Serum of Rats with Aplastic Anemia on Mechanism of Proliferation and Differentiation of Hematopoietic Progenitor Cells / 世界科学技术-中医药现代化

This study was aimed to investigate the effect of Bu-Shen Y i-Sui Sheng-Xue (BSYSSX) method on pro-liferation and differentiation mechanisms of hematopoietic progenitor cells. The rat models were established by 60Co-γrays and cyclophosphamide. Compound Chinese medicine was gavaged to rats of the normal control group, model group, stanozolol group, Yi-Sui Sheng-Xue (YSSX) group, Wen-Shen Sheng-Xue(WSSX) group and Zi-Shen Sheng-Xue (ZSSX) group. Then, serum of rat was prepared. Rat bone marrow cells were incubated with AA rats serum ac-counted for 20% and the number of hematopoietic progenitor cells colony-forming units (CFU) were counted. The level of GATA-1 and PU.1 mRNA in colony cells were detected with RT-PCR. The results showed that compared with the normal control group, the number of bone marrow cells, CFU-E, BFU-E, CFU-GM, as well as the expres-sion of GATA-1 and PU.1 mRNA in the model group decreased significantly (P< 0.01). Compared with the model group, the number of bone marrow cells, CFU-E, BFU-E, CFU-GM of each treatment group were significantly in-creased (P< 0.01). CFU-E and BFU-E of the ZSSX group were better than the YSSX group (P < 0.01). CFU-GM of the ZSSX group was better than the YSSX group and the WSSX group. The expression of GATA-1 and PU.1 mR-NA in each treatment group were significantly higher than the model group (P< 0.01). The expression of GATA-1 mRNA in the ZSSX group was better than the YSSZ group and WSSX group (P< 0.05). The expression of PU.1 mR-NA in the ZSSX group was higher than the YSSX group and WSSX group. It was concluded that BSYSSX method may increase the expression of GATA-1 and PU.1 mRNA in order to promote the proliferation and differentiation of bone marrow hematopoietic progenitor cells. The ZSSX method was better than the YSSX method and WSSX method.

Effect of different doses of HS6101 on hematopoietic function of ICR mice treated with cyclophosphamide / 国际药学研究杂志

Objective To observe the effect of different doses of HS6101 on the recovery of hematopoietic injury in ICR mice treated with cyclophosphamide (CTX). Methods Normal ICR mice were intraperitoneally injected with CTX at 100 mg/kg once a day for 3 consecutive days,and the mouse model of chemotherapy-induced hematopoietic injury was established. Three groups of mice (with 20 per group),were respectively injected with HS6101 at 0,9 or 27 μg subcutaneously at one hour before the first administration of CTX. The peripheral blood cell counts of the mice were observed once every 2 days. Hematopoietic progenitor cell colony counting and histopathological assessment of bone marrow cells were evaluated at 4 d and 9 d after the first administration of CTX. Results In ICR mice after chemotherapy with CTX,all doses of HS6101 significantly increased peripheral leukocytes and neutrophils (P<0.05),elevated the number of multilineage hematopoietic progenitor cell colonies of bone marrow,and stimulated the proliferation of bone marrow cells after CTX injury. Mice receiving 27 μg HS6101 were better than those of the other two groups. Conclusion HS6101 at 27 μg could significantly promote the recovery of hematopoiesis in ICR mice treated with CTX chemotherapy.

A infecção das células progenitoras hematopoiéticas pelo HIV e a relação com a persistência viral e anormalidades hematológicas / Infection of hematopoietic progenitor cells by HIV and the relation with viral persistence and hematological abnormalities

O vírus da imunodeficiência humana causa infecção persistente apesar da terapia com antirretrovirais. Isso leva a crer que existem outros reservatórios virais além dos já conhecidos. Estudos recentes mostram que o vírus pode invadir as células progenitoras hematopoiéticas, causar infecção ativa assim como latente e morte celular. Essas célulasinfectadas latentemente podem persistir por vários anos e contribuir para a viremia residual e a persistência viral. A morte celular causada pela invasão e replicação do vírus pode estar associada a anormalidades hematopoiéticas. Palavras-chave. Células progenitoras hematopoiéticas; vírus da imunodeficiência adquirida; reservatórios virais

Human immunodeficiency virus (HIV) causes persistent infection despite antiretroviral therapy. This suggests the existence of other viral reservoirs beyond those already known. Recent studies show that HIV can invade hematopoietic progenitor cells, generating active infection as well as latent infection and cell death. The latently infected HPCs may persist for several years and contribute to residual viremia and viral persistence. Cell death caused by the invasion and replication of the virus may be associated with hematopoietic abnormalities.

Evaluación de la contaminación microbiológica en productos de células progenitoras hematopoyéticas. Experiencia de un laboratorio de procesamiento de CPH y Banco de Sangre de Cordón Umbilical / Microbiological evaluation in hematopoietic progenitor cell products. Experience from an HPC processing laboratory and Cord Blood Bank

Los productos de células progenitoras hematopoyéticas (CPH) representan una fuente potencial de infección en pacientes inmunosuprimidos que reciben infusión de CPH como parte de su tratamiento. La probabilidad de contaminación de cada producto difiere según la técnica de colecta y el procesamiento aplicado. En este trabajo hemos realizado un análisis retrospectivo de los resultados de cultivos microbiológicos de 1707 productos de CPH obtenidos de sus tres fuentes (médula ósea, sangre periférica y sangre de cordón umbilical) con el objetivo de determinar la proporción de unidades contaminadas. Además, fueron comparadas las distintas técnicas de colecta y las diferentes manipulaciones a las que fueron sometidos los productos. Por otro lado, se analizaron las posibles fuentes de contaminación según el microorganismo identificado y se evaluó la supervivencia de los mismos luego del descongelamiento. La prevalencia de productos de CPH con cultivos microbiológicos positivos reportados en este estudio (5,2%) se corresponde con lo descripto en la literatura. No encontramos diferencias significativas al comparar los productos según la fuente de la cual provenían las CPH. Tampoco hubo diferencias según los procedimientos aplicados a cada unidad. Los microorganismos aislados en los productos de CPH fueron los esperados de acuerdo a la fuente de la cual provenían las células. Pudo comprobarse que algunos de ellos son capaces de sobrevivir a los procesos de criopreservación y descongelamiento. La estricta adhesión a las normas de buenas prácticas de manufactura y buenas prácticas tisulares es un requisito para minimizar los riesgos de introducir microorganismos contaminantes. Disponer de un producto de CPH seguro es fundamental para el éxito de un trasplante.

Hematopoietic stem cell products (HSCP) represent a potential source of infection for immunosuprressed patients that receive HSCP infusion as part of their treatment. The probability of contamination of a HSCP product depends on the collection technique as well as the processing performed. In this work we have carried out a retrospective analysis of the results of 1707 microbiological cultures of HSCP products obtained from three different sources: bone marrow, peripheral blood and umbilical cord blood. We determined the proportion of HSCP units that were contaminated by microorganisms. Furthermore we compared the results obtained with different collection techniques and with the distinct manipulations that were used for processing. Moreover we analysed the possible sources of contamination related to the microorganisms identified and we evaluated the survival of them after thawing. The proportion of HSCP products with positive microbiological cultures obtained in this study (5,2%) correlates with that reported by other authors. We have not found significant differences between the results achieved with HSCP products from different sources. There were neither differences depending on the procedures applied. The isolated microorganisms from the HSCP products were the expected in accordance with the source of the cells. It could be demonstrated that some of them were capable of surviving the cryopreservation and thawing processes. Adherence to good manufacture practices and good tissue practices regulations is critical for minimizing the risks of introducing contaminant microorganisms. A safe HSCP product is essential for the success of a transplant.

Inverse Tendency between Ex Vivo Expansion Potential of Hematopoietic Progenitors and Time to Engraftment after Hematopoietic Stem Cell Transplantation / 대한진단검사의학회지

BACKGROUND: The CD34+ cell dose and infused number of committed progenitor cells in transplantation are important factors in hematologic engraftment. However, the relationship between expansion potential of progenitor cells and hematologic engraftment remains controversial. We evaluated whether expansion potential of progenitor cells is a predictive factor of post-transplantation hematologic engraftment. METHODS: Mononuclear cells isolated from mobilized peripheral blood and bone marrow were cultured with cytokine cocktail for 7 days. Progenitor cells and committed progenitors were analyzed using stem cell markers (CD34 and CD133) and lineage specific markers. Hematologic engraftment was defined as neutrophil counts over 500/microliter and platelet counts over 20,000/microliter without transfusion. Acute and chronic graft-versus-host disease (GVHD) were investigated. RESULTS: There was inverse tendency between the number and fold expansion of progenitor cells or committed (granulocytic or megakaryocytic) progenitors and time to engraftment. Especially, fold expansion of CD34(+)/CD33(+) cells was significantly correlated with time to neutrophil engraftment in bone marrow transplantation (r=-0.56, P=0.04). The infused number and fold expansion of lymphoid progenitors were not related to the occurrence of acute or chronic GVHD. CONCLUSIONS: We could not prove that expansion potential of progenitor cells and committed progenitor cells is correlated to hematologic engraftment although there is a correlation between CD34(+)/ CD33(+) cells and time to neutrophil engraftment. But, a further study on the value of expansion potential is required because there is an inverse tendency.

The Expression of Fas antigen and Bax and Apoptosis in Ex Vivo Expanded Hematopoietic Progenitor Cells / 대한혈액학회지

BACKGROUND: During ex vivo expansion of cord blood (CB) CD34+ cells, differentiation of the expanded cells happened and hematopoietic potential of the progenitor cells decreased. In this study, we evaluate the effect of the expression of Fas antigen, Bcl-2, and Bax on CD34+ or AC133+ hematopoietic progenitor cells during ex vivo expansion. METHODS: CD34+ and AC133+ cells isolated from human CB were cultured in serum free medium supplemented with several cytokines for 7 days. After expansion culture, we re isolated CD34+ and AC133+ cells and compared the numbers of granulocyte-macrophage colony-forming units (CFU-GM) and granulocyte, erythrocyte, monocyte, and macrophage colony-forming units (CFU-GEMM), and expression of Fas antigen, Bcl-2, and Bax with unexpanded cells. RESULTS: CFU-GM was expanded 23.94 fold in CD34+ cells and 15.22 fold in AC133+ cells at day 7 of culture but CFU-GEMM was not expanded. The expression of Fas antigen and Bax was 7.44% and 2.75%, respectively, in fresh isolated CD34+ cells and increased to 19.71 % and 33.67%, respectively, in expanded CD34+ cells at day 7 culture, but Bcl-2 was not changed. In case of AC133+ cells, the expression of Fas antigen and Bax were also increased from 5.87% and 6.19% to 24.85% and 22.83%, respectively, and Bcl-2 was slightly decreased. Apoptosis was not changed in CD34+ cells and AC133+ cells during ex vivo expansion. CONCLUSION: These results indicate that the nature of expansion was similar between CD34+ and AC133+ cells, and expression of Fas antigen and Bax increased on CD34+ and AC133+ cells during ex vivo expansion. Selection of the expanded progenitor cells without apoptosis may be useful for the hematopoietic stem cell transplantation.

Ex vivo Expansion of Hematopoietic Progenitor Cells in Co-culture of Cord Blood CD34+Cells with Human Umbilical Vein Endothelial Cells / 대한혈액학회지

BACKGROUND: We examined an ex vivo expansion system for cord blood (CB) hematopoietic progenitor cells, which is based upon a co-culture of CD34+cells with human umbilical endothelial cells (HUVECs) in the presence of stromal cell-derived factor-1 (SDF-1) and hematopoietic growth factors. METHODS: Cord blood CD34+cells were either incubated a liquid suspension culture or co-cultured on HUVEC monolayers with hematopoietic growth factors in the presence or absence of SDF-1. After 7~14 days of culture, cells were harvested and analyzed for fold-increase in nucleated cells, CD34+ cells, and colony-forming cells (CFCs) and apoptosis. RESULTS: Seven-day suspension culture of CD34+ cells in the presence of a cytokine combination consisting of throbmopoietin, flk-2 ligand, and kit-ligand (TFK) led to a 43-fold increase of nucleated cells, a 19-fold increase of CD34+ cells, and 14-fold increase of CFCs, respectively. The addition of SDF-1 to TFK slightly further increased this expansion. A co-culture of CD34+ cells with HUVECs significantly enhanced the expansion of both CD34+ cells and CFCs compared with a liquid suspension culture. This was further increased by the addition of SDF-1. A co-culture of CD34+ cells on HUVECs transfected with null-adenoviral vector led to a better fold increase of haemtopoietic progenitor cells compared with a culture with non-transfected HUVECs. Adding SDF-1 to the co-culture diminished the annexn V-positive cells both in the supernatant and adherent cell layers. CONCLUSION: A co-culture of cord blood cells with HUVECs in the presence of hematopoietic growth factors and SDF-1 could be a new model for the efficient expansion of hematopoietic progenitors.

Study of the effect of iron overload on the bone marrow hematopoietic function in immuo-related pancytopenia patients / 中国实用内科杂志

Objective To study the effect of iron overload on the bone marrow hematopoietic function on immuo-related pancytopenia(IRP)patients by hematopoietic progenitor cell(HPC)culture of bone marrow(BM).Methods BM liquid 4～5 mL was taken from 46 IRP patients of General Hospital Tianjin Medical University from July 2009 to February 2010 to detect colony-forming-unit of erythrocyte,blast-forming-unit of erythrocyte and colony-forming-unit of granulocyte-monocyteby HPC culture of BM.And according to the serum ferritin(SF)level,these patients were classified into 2 groups to compare HPC proliferation,blood cell counts,BM proliferation,blood transfusion and treatment effects.Results The mean values of 3 different colonies of IRP patients with high SF level [(43.33?17.74),(1.50?2.2),(11.06?5.83)/105BMMNC] were significantly lower than those of the patients with normal SF level [(77.43?40.64),(9.57?7.99),(21.25?11.41)/105BMMNC](P

Committed differentiation of CD34~+ hematopoietic progenitor cells / 中国癌症杂志

Purpose:To study the capacity of hematopoietic growth factors combinations(IL 3,G CSF,GM CSF) for inducing CD34 + cells to proliferate and differentiate committedly to granulocytes. Methods:CD34 + cells were isolated from peripheral blood by using a high gradient magnetic cell sorting sysem(MACS),and expanded with IL 3、G CSF and GM CSF in a liquid culture system.Colony forming cells and CD34 + cell ratio were studied by colony forming assays and FACS. Results:The combinations of IL 3,G CSF and GM CSF increased the total cell number by 56 fold and 110 fold, increased CFU GM by 8.44 fold and 35.08 fold respectively at the 3rd and the 7th day after culturing ,and the ratio of CD34 +cell decreased during the culture.Conclusions:It is possible to induce CD34 +cells to proliferate and differentiate committedly to granulocyte lineage.

Effects of rhIL-17 on Differentiation and Development of Murine Hematopoietic Progenitors and Human Cord Blood CD34+ Stem Cells / 中国肿瘤生物治疗杂志

To investigate effects of rhIL-17 on growth and development of mouse bone marrow progenitors andhuman cord blood LD34~+ stem cells. Methods: Mouse bone marrow progenitors were isolated by routine protocol, and CD34~+ stem cells were isolated from normal human cord blood by Mini-MACS, then cultured with rhIL-17 and/or GM-CSF/IL-4. The phenotypes of the cells were analyzed by FACS, IL-12 level was analyzed by ELISA and T cell stimulating activity in allo-MLR was determined by [~3H]-TdR incorporation. Results: Expression of MHC class II molecules and B7-2 on the surface of immature DC derived from mouse bone marrow progenitors was up-regulated by IL-17. The capacity of the cells to secrete IL-12 and their T cell stimulating activity were also enhanced. The cells showed the characteristics of mature DC. After cultured with IL-17 for 9 days, the number of CD34~+ stem cells increased by 2 times. The phenotypes of some cells were CDla~(high), B7-2~(high), and HLA-DR~(lwo). The cells could stimulate allo geneic T cells to proliferate but their capacity was lower than that of the cells cultured with IL-17 combined with GM-CSF. The cells cultured with IL-17 and GM-CSF proliferated markedly and the rate of CDla~+ and B7-2~+ cells increased significantly. The T cell stimulating activity of cells was also augmented. Conclusion: IL-17 could promote DC derived from mouse bone marrow progenitors to mature. When combined with GM-CSF, IL-17 could induce human CD34~+ stem cells not only to proliferate markedly but also to show characteristics of DC, indicating that CD34~+ stem cells might differentiate to DC by IL-17.

Effect of angelica polysaccharide on bone marrow macrophage and its relationship to hematopoietic regulation / 中草药

Objective To investigate the effect of angelica polysaccharide (APS) on bioactivity of bone marrow macrophage (BMM?) and its relationship to hematopoietic regulation, for clarifying the hematonic mechanism of Angelica sinensis. Methods The techniques of hematopoietic progenitor cell culture and BMM? culture in vitro, biological assay of hematopoietic growth factor (HGF) in culture media of BMM?, immunocytochemistry, and nucleic acid in situ hybridization were used. Results The culture supernatant of BMM? induced by APS can enhance the CFU-Mix, CFU-E, CFU-GM; the expression of erythropoietin (EPO), GM-CSF, IL-3, IL-6 protein in BMM? induced by APS was much stronger than that in the control group at different levels; the expression of EPO mRNA and GM-CSF mRNA in BMM? induced by APS were intensified. Conclusion APS may directly and/or indirectly stimulate the BMM? in hematopoietic inductive microenvironment to accelerate the synthesis and secretion of hematopoietic regulation factors on the basis of gene and protein level, such as EPO, GM-CSF, IL-3, IL-6, which in turn to promote the proliferation and differentiation of CFU-Mix, CFU-E, and CFU-GM.

EFFECT OF RECOMBINANT HUMAN INTERLECUKIN-11(rhIL-11)ON PERIPHERAL PLATELET COUNT AND HEMATOPOIETIC PROGENITOR PROLIFERATION / 解放军医学杂志

In order to obtain experimental evidences of therapeutic effect of rhIL 11 for clinical use, we observed the effects of rhIL 11 on colony formation of multilineage human hematopoietic cells in vitro and acceleration of platelet count and bone marrow hematopoietic recovery in 3 0Gy ? ray irradiated rhesus monkeys. The results showed that rhIL 11 significantly increased the nadirs of PLT count and shortened the duration of PLT numbers below 50% of their baseline values. The recovery of PLT in irradiated monkeys was accelerated. Dosage related PLT recovery was not observed. rhIL 11 not only promoted the proliferation and maturation of CFU Mk but also improved the proliferation of CFU GM, CFU e, BFU e and CFU Mix. These data suggest that rhIL 11 may be an effective agent in the treatment of hemopoietic suppression and thrombocytopenia.